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Method for isolating apoplast effector protein secreted by pathogenic bacteria from plant tissues infected with pathogenic bacteria

A technology for plant tissue and pathogenic bacteria is applied in the field of isolating apoplast effector proteins secreted by pathogenic bacteria to avoid pollution and ensure no damage.

Active Publication Date: 2021-06-04
北京中农探味科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, there is no relevant report in the prior art about isolating and extracting the apoplast juice containing the effector protein secreted by pathogenic bacteria from plant tissues infected with pathogenic bacteria

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Isolating the apoplast juice containing the apoplast effector protein secreted by the Fusarium verticillium from corn plants infected with the Fusarium verticillium comprises the following steps:

[0065] (1) Take the stem node of the corn plant infected with Verticillium spp., place it in a beaker equipped with 25mM Tris, 100mM NaCl (PH=7.4) buffer solution and soak for 60s;

[0066] (2) Take out the corn plant stem node after soaking, blot the surface buffer with absorbent paper, and cut into thin slices about 10mm thick with a sterilized scalpel blade;

[0067] (3) the corn plant stem node after sectioning is weighed for the first time, and record the first quality;

[0068] (4) Put the sliced ​​corn plant stem nodes into a 50mL syringe, add buffer until the corn plant stem nodes are completely submerged, discharge excess air in the syringe barrel, seal the needle port of the syringe, and pull the plunger of the syringe to 50mL of the syringe Then release the plunge...

Embodiment 2

[0074] Isolating the apoplast juice containing the apoplast effector protein secreted by the Fusarium verticillium from corn plants infected with the Fusarium verticillium comprises the following steps:

[0075] (1) Take the stem nodes of corn plants infected with Fusarium verticillium, place them in a beaker equipped with 100mM NaCL (PH=7.4) buffer solution and soak for 30s;

[0076] (2) Take out the corn plant stem node after soaking, blot the surface moisture with absorbent paper, and cut into thin slices about 5mm thick with a sterilized scalpel blade;

[0077] (3) the corn plant stem node after sectioning is weighed for the first time, and record the first quality;

[0078] (4) Put the sliced ​​corn plant stem nodes into a 50mL syringe, add buffer until the corn plant stem nodes are completely submerged, discharge excess air in the syringe barrel, seal the needle port of the syringe, and pull the syringe plunger to the syringe 50mL scale, and then release the piston at a...

Embodiment 3

[0084] Isolating the apoplast juice containing the apoplast effector protein secreted by the Fusarium verticillium from corn plants infected with the Fusarium verticillium comprises the following steps:

[0085] (1) get the stem node that corn plant is infected with Verticillium spp., place in the beaker that 25mM Tris buffer solution is housed and soak for 50s;

[0086] (2) Take out the corn plant stem node after soaking, blot the surface buffer with absorbent paper, and cut into thin slices about 15mm thick with a sterilized scalpel blade;

[0087] (3) the corn plant stem node after sectioning is weighed for the first time, and record the first quality;

[0088] (4) Put the sliced ​​corn plant stem nodes into a 50mL syringe, add buffer until the corn plant stem nodes are completely submerged, discharge excess air in the syringe barrel, seal the needle port of the syringe, and pull the syringe plunger to the syringe 50mL Then release the plunger at a constant speed of 60mL / m...

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Abstract

The present disclosure relates to a method for isolating apoplast juice from plant tissues infected with pathogenic bacteria, said apoplast juice containing effector proteins secreted by said pathogenic bacteria, the method comprising: a, placing plant tissues infected with pathogenic bacteria in Carry out infiltration treatment under vacuum in the buffer solution to obtain the plant tissue after the infiltration treatment; b. Centrifuge the plant tissue after the infiltration treatment for the first time to obtain the first centrifugate; take the first centrifugation solution for the second Sub-centrifugation to collect the second centrifugate; wherein, in the plant tissue infected with pathogenic bacteria, the pathogenic bacteria are Fusarium graminearum, Fusarium verticillium, Fusarium oxysporum, Verticillium dahliae, Anthracnose graminearum , Magnaporthe oryzae, Botrytis cinerea at least one. This method can maximize the separation of the effector proteins secreted by pathogenic bacteria and enter into the apoplast juice, which facilitates better analysis of the components and characteristics of the apoplast effector proteins secreted by pathogenic bacteria in susceptible plant tissues.

Description

technical field [0001] The present disclosure relates to the technical field of extraction and separation of pathogenic bacteria effector proteins, in particular to a method for isolating apoplast effector proteins secreted by pathogenic bacteria from plant tissues infected with pathogenic bacteria. Background technique [0002] Plant apoplast refers to the system composed of cell wall, intercellular space and vessel on the periphery of plant cell protoplast. Many important biological processes occur in the apoplast juice, such as cell wall formation, nutrient uptake and transport, plant-pathogen interaction, etc. For plants infected with pathogenic bacteria, the apoplast is the core of the interaction between the pathogen and the host, the pathogen will secrete apoplast effector protein to inhibit the innate immunity of the host, and the host will also secrete corresponding antibacterial substances to inhibit the infection of the pathogen . [0003] The existing method fo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K1/14
CPCC07K1/145
Inventor 郭维张君席凯飞
Owner 北京中农探味科技有限公司
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